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Related Concept Videos

The Evidence for Evolution02:55

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Genetic variations accumulating within populations over generations give rise to biological evolution. Evolutionary changes can result in the formation of novel varieties and entire new species. These changes are responsible for the diverse forms of life inhabiting the planet. The evidence for evolution suggests that all living organisms descended from common ancestors.
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The endosymbiont theory is the most widely accepted theory of eukaryotic evolution; however, its progression is still somewhat debated. According to the nucleus-first hypothesis, the ancestral prokaryote first evolved a membrane to enclose DNA and form the nucleus. Conversely, the mitochondria-first hypothesis suggests that the nucleus was formed after endosymbiosis of mitochondria.
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The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
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Fluorescence Activated Cell Sorting of Plant Protoplasts
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Fluorescence-Activated Droplet Sorting for Single-Cell Directed Evolution.

Derek Vallejo, Ali Nikoomanzar, Brian M Paegel1

  • 1Department of Chemistry , The Scripps Research Institute , Jupiter , Florida 33458 , United States.

ACS Synthetic Biology
|May 24, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed a new fluorescence-activated droplet sorting (FADS) instrument to evolve enzymes for artificial genetic polymers. This directed evolution method enhances enzyme activity for synthetic biology applications, improving health and environmental solutions.

Keywords:
DrOPSdroplet microfluidicsdroplet sortingenzyme engineeringhigh throughput screening

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Area of Science:

  • Synthetic biology
  • Biotechnology
  • Enzyme engineering

Background:

  • Natural enzymes have suboptimal activity in engineered pathways or with unnatural substrates.
  • Directed evolution is crucial for discovering new enzyme variants with desired functions.
  • Artificial genetic polymers (XNAs) offer novel applications but require tailored enzymes.

Purpose of the Study:

  • To develop and validate a fluorescence-activated droplet sorting (FADS) instrument.
  • To enable efficient directed evolution of enzymes for XNA synthesis and modification.
  • To advance synthetic biology tools for practical applications.

Main Methods:

  • Construction and validation of a microfluidic FADS instrument.
  • Utilized fluorescent sensors responsive to enzymatic activity for droplet sorting.
  • Applied the FADS system for directed evolution of enzymes targeting XNAs.

Main Results:

  • The FADS instrument enables droplet sorting at high throughput (∼2-3 kHz).
  • Demonstrated successful evolution of enzymes for synthesizing and modifying XNAs.
  • The system effectively identifies and selects enzyme variants with improved activity.

Conclusions:

  • The developed FADS instrument is a powerful tool for enzyme evolution in synthetic biology.
  • This technology facilitates the customization of enzymes for novel applications, including XNA technologies.
  • Advances in enzyme engineering through FADS will drive innovation in biotechnology and healthcare.